Fast liquid chromatography coupled to electrospray tandem mass spectrometry peptide sequencing for cross-species protein identification

Authors

  • Erwin Witters,

    Corresponding author
    1. Universiteit Antwerpen, UIA, dept. Biologie, Laboratorium voor plantenbiochemie, Universiteitsplein 1, B-2610 Antwerpen, Belgium
    • Universiteit Antwerpen, UIA, dept. Biologie, Laboratorium voor plantenbiochemie, Universiteitsplein 1, B-2610 Antwerpen, Belgium.
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  • Kris Laukens,

    1. Universiteit Antwerpen, UIA, dept. Biologie, Laboratorium voor plantenbiochemie, Universiteitsplein 1, B-2610 Antwerpen, Belgium
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  • Peter Deckers,

    1. Universiteit Antwerpen, UIA, dept. Biologie, Laboratorium voor plantenbiochemie, Universiteitsplein 1, B-2610 Antwerpen, Belgium
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  • Walter Van Dongen,

    1. Universiteit Antwerpen, RUCA, dept. Scheikunde, Nucleoside Research and Mass Spectrometry Unit, Groenenborgerlaan 171, B-2020 Antwerpen, Belgium
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  • Eddy Esmans,

    1. Universiteit Antwerpen, RUCA, dept. Scheikunde, Nucleoside Research and Mass Spectrometry Unit, Groenenborgerlaan 171, B-2020 Antwerpen, Belgium
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  • Harry Van Onckelen

    1. Universiteit Antwerpen, UIA, dept. Biologie, Laboratorium voor plantenbiochemie, Universiteitsplein 1, B-2610 Antwerpen, Belgium
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Abstract

Using a parallel microcolumn switching liquid chromatography set-up coupled to a quadrupole time-of-flight mass spectrometer, a rapid liquid chromatography/mass spectrometric (LC/MS) protein identification method is presented. Without prior sample clean-up up to 300 protein digest samples a day can be processed. Using data-directed acquisition, up to 10 fragmentation analyses for each protein sample can be acquired in the same chromatographic run that can be used for database searching. Using internal peptide sequence information, protein databases and the various nucleic acid databases can both be queried for cross-species identification of the protein sample. The method was evaluated and put into force to generate data for a tobacco cell culture protein database. Copyright © 2003 John Wiley & Sons, Ltd.

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